Field of Invention
The present invention relates to the field of molecular biology, specifically to a human rare blood group detection method, kit, rapid screening method and application.
Description of Related Arts
Rare blood group antigen is a blood group antigen whose frequency in the public is less than one-thousandth, and due to its scarcity, in blood transfusion of patients of a rare blood group, it is difficult to find the source of matched blood, resulting in delay of treatment. Thus, rare blood group detection, rapid screening and establishment of a rare blood group bank are extremely important.
The existing rare blood group detection method mainly includes:
1. Serological Methods
(1) Use human polyclonal sera, take advantage of the saline agglutination test or indirect antiglobulin test to perform rare blood group detection by a tube method, a U-shaped 96-well microtiter plate method or a gel method.
(2) Use animal origin (mainly murine) monoclonal or polyclonal antibody reagents of IgM-type or IgG-type, take advantage of the saline agglutination test or indirect antiglobulin test to perform rare blood group detection by a tube method, a U-shaped 96-well microtiter plate method or a gel method.
(3) For special Jk(a−b−) phenotype, a Urea method can be used. Jk(a−b−) phenotype red blood cell (RBC) may maintain the membrane integrity for 30 minutes in a 2M urea solution, and whether to be Jk(a−b−) phenotype is judged according to whether a subject RBC is hemolysis in the 2M urea solution after 10 minutes.
2. Molecular Biology Methods: Design Based on the RBC Antigen SNPs
(1) Middle and low-throughput genotyping: including PCR-RFLP, PCR-SSP, real-time quantitative PCR, and pyrosequencing techniques.
(2) High-throughput genotyping: including commercial or non-commercial Beadchip, BloodChip, GenomeLab SNP stream, Luminex xMAP, and solid-phase hybridization techniques.
3. Review: the rare blood group samples obtained through screening with the above two methods generally should be reviewed through serological methods such as saline agglutination test or indirect antiglobulin test.
The existing rare blood group detection method has the following problems:
1) Serological methods: it is difficult for the majority of RBC rare blood group antigens to obtain human antibody or commercialization detection reagent, or as reagent price is too high and screening is costly, it is difficult to achieve large-scale high-throughput screening.
2) Molecular biology methods: middle and low-throughput genotyping methods are often time-consuming, and thus it is difficult to achieve large-scale high-throughput screening. The existing high-throughput genotyping methods are expensive. And the existing genotyping methods detect a single specimen, and detection costs will undoubtedly increase.
Therefore, finding a fast, economical, efficient rare blood group detection and screening method is of great practical significance.